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R Müllenbach

Publications and source records attributed to R Müllenbach.

13 recordsLinked to original sources
PubMed1997

A 5.5-Mb high-resolution integrated map of distal 11q13.

The distal part of 11q13, which contains several genes relevant to human diseases, has been poorly mapped as part of genome-wide mapping efforts. In the prospect of drawing a fine-scale integrated map of the area containing KRN1 and OMP, we have established a framework of markers by hybridization to DNA of somatic cell hybrids and by fluorescence in situ hybridization (FISH) on metaphase chromosomes. The probes studied were used to isolate 27 YACs and 16 cosmids that could be organized in three contigs covering approximately 6 Mb. These contigs were separated by two gaps that are likely to contain sequences underrepresented in YAC libraries. They were then integrated based on long-range restriction mapping and DNA-fiber FISH into a high-resolution physical map, which covers a 5.5-Mb region and includes 36 anonymous markers and 10 genes. This map will be used to search for genes within the 2/3 of this region where none have been localized as yet. It will also lay the ground for the characterization of an amplicon surrounding GARP in breast cancer and for the search of disease genes within this region.

Animals↗
PubMed1996

Cosmid-derived transcripts and sequence tags mapped to three subregions of human chromosome 22.

Fifty cosmids from the ICRF, London, and Lawrence Livermore Laboratory, California, human chromosome 22 cosmid libraries were isolated, regionally assigned and tested for their ability to detect repeats or single copy sequences. The search resulted in nine cosmids containing repetitive motifs from the pericentric region of chromosome 22. An additional 19 cosmids, that detected single copy sequences in the long arm of chromosome 22q: 7 in the region 22q11.2-q13.1 and 12 in 22q13.1-qter, were mapped more precisely by fluorescence in situ hybridization. Three out of these 19 recombinants displayed restriction fragments containing (CA)n repeats, were subcloned and sequenced. One cosmid, representing a region coding for an ubiquitous 300-bp transcript, is localized 600 kb from PDGFB, and four cosmids contained sequences surrounding the ARSA gene at 22q13.3. Presently, long range physical maps, that may be useful for analysing structural alterations of chromosome 22q13, are being constructed from these additional, regionally assigned markers from chromosome 22q13 employing both existing cosmid and new bacterial artificial chromosome (BAC) libraries.

Chromosome Mapping↗
PubMed1996

Distribution and linkage of repetitive clusters from the heterochromatic region of human chromosome 22.

The pericentric regions of eukaryotic chromosomes consist of several types of repetitive DNA families. In human chromosome 22, the organization of such families was studied in more detail. In addition to the known families of alpha and beta repeats, an additional repeat with a 48-bp motif was previously assigned to 22pter-q11. Here, we report in more detail the distribution of these repeat families, applying pulsed-field gel electrophoresis, fluorescence in situ hybridization and physical linkage on cosmid recombinants. At least two clusters of 48-bp repeats are localized on chromosome 22, one on the distal p-arm and one in the region 22cen-q11. Cosmids from a chromosome 22 library, containing both 48-bp and beta-repeats, link both arrays on 22p and define their maximum distances to less than 44 kb. Loss of 48-bp repeat sequences in a Dl-George cell line carrying a deletion in 22q11 suggests the presence of a second cluster in 22q11, a distribution supported by (fluorescene in situ hybridization)-FISH signal analysis. As additional members of the 48-bp repeat family can be found on all acrocentric chromosomes. It remains to be determined whether the distribution seen on chromosome 22 is also common in other human acrocentric chromosomes.

Centromere↗
PubMed1994

Debrisoquine hydroxylase gene polymorphism in meningioma.

Cytochrome P450 CYP2D6 polymorphism is an autosomal recessive trait associated with impaired debrisoquine metabolism in 5-10% of caucasian populations. This polymorphism has been associated with susceptibility to Parkinson's disease, bladder cancer, various forms of leukemia and possibly melanoma. In many other cancer forms, the data remained contradictory due to the technical limitations for identifying affected individuals (poor metabolizers). A recently developed polymerase chain reaction-based assay allows convenient screening of approximately 80% of known mutations. We have tested brain tumors correlated with chromosome 22 deviations for genetic polymorphism in the cytochrome P450 CYP2D6 locus localized on chromosome 22q13. Thirty-one meningioma samples were analyzed and the observed frequency of heterozygotes and homozygotes for the G to A mutation did not deviate significantly from the distribution in a normal population. These data are comparable to previous observations in for example breast and colon cancer and indicate that the CYP2D6 locus on chromosome 22q13 is not involved in the pathogenesis of meningiomas.

Alleles↗
PubMed1994

PCR expression analysis of the estrogen-inducible gene BCEI in gastrointestinal and other human tumors.

A polymerase chain reaction (PCR) assay was developed to test for tumor cell specific expression of the BCEI gene. This new marker gene, reported at first for human breast cancer, was found specifically active in various gastrointestinal carcinomas by previously applying immunohistochemistry and RNA (Northern blot) analysis. Presently, by using reverse transcription-PCR analysis, a series of primary tumor tissues and established tumor cell lines were tested for BCEI transcription. This approach was compared to immunostaining achieved by an antibody directed against the BCEI gene's product. The result demonstrate the superior sensitivity of PCR by indicating the gene's expression in cases where immunohistochemical testing remained negative.

Base Sequence↗
PubMed1994

Sequence patterns and hybridization analysis of clones generated by Alu-PCR.

A series of clones from an Alu-PCR library were analysed in more detail. Characterization by Southern blot hybridization and sequencing displayed several features common to all probes generated by this approach: Short length of the PCR-products as well as the presence of homologous regions on both ends resulted in a limited feasibility for filter hybridization and a low probability of restriction length polymorphisms. In addition, a series of different short repeats at the 3'-ends of Alu-repeats and present in the generated probes offers a rich source of potential variable sites accessible by PCR.

Animals↗
PubMed1993

Cosmid mapping and locus linkage within the human chromosomal region 22q13.1.

Many human meningiomas show loss of heterozygosity at distal loci but retain constitutional heterozygosity at one or more proximal loci of 22q. Molecular analysis indicted deletions involving at least the region 22q12.3-qter. In this region, distal to myoglobin, the putative meningioma locus ought to be expected. Long-range mapping was performed around two loci from 22q12.3-q13.1 (D22S16 and PDGFB, the most proximal locus to be lost in meningioma). D22S16, originally assigned to 22q13-qter by isotopic in situ hybridization, was placed in the vicinity of PDGFB by utilizing a set of somatic cell hybrids, an assignment confirmed by fluorescence in situ hybridization (FISH) of a cosmid clone containing the D22S16 locus. Moreover, pulsed field gel electrophoresis suggests a close linkage of both markers within 630 kb.

Arachnoid↗
PubMed1993

Application of DNA filter hybridization and PCR to distinguish between human and non-human tissues of poor quality.

The present report describes a novel approach for the identification of human or non-human specimens after long-term storage in a badly preserved state. The application of the PCR-technique (polymerase chain reaction) using human-specific primers as well as Southern blot (filter) hybridization of the sample DNA to a primate-specific DNA probe enabled us to extend the positive identification beyond the limits of conventional methods such as serological or morphological examinations.

Animals↗
PubMed1992

A non-alphoid repetitive DNA sequence from human chromosome 21.

A non-alphoid repetitive DNA from human chromosome 22, consisting of a 48-bp motif, shows homology to both G-group chromosomes in the gorilla, thus indicating the presence of additional repeat family members on further human chromosomes. Therefore, we screened a chromosome-21-specific cosmid library using this repetitive sequence from chromosome 22 (D22Z3). Some 40-50 cosmid clones were positive in tests for hybridization. One of the clones giving the strongest signals was digested with EcoRI/PstI, which we knew to cut frequently within the repeats; this resulted in fragments containing repeat units only. The fragments were subcloned into plasmid vector pTZ 19. Sequence-analysis of a 500-bp insert showed ten copies of a 48-bp repeat similar to D22Z3, with about 15% sequence deviation from the chromosome 22 consensus sequence. In situ hybridization of the newly isolated recombinant established its chromosome 21 specificity at high stringency. Physical mapping by pulsed field gel electrophoresis placed this new repeat in close vicinity to the chromosome 21 alphoid repeat. No cross-hybridization with other mammalian genomes except for those of apes was observed. The locus has been designated D21Z2 by the Genome Data Base. A gel mobility shift assay indicated that this repetitive motif has protein-binding properties.

Animals↗